Objective To observe the effects of olfactory lamina propria (OLP) transplantation and ganglioside GM1 treatment on spinal cord injury (SCI) in rats.Methods Totally 50 healthy pure breed female Sprague-Dawley (SD) rats after spinal cord hemiseetion were randomly divided into 5 groups and were given different treatments: (OLP + GM1) treatment group (group A), GM1 treatment group (group B), OLP treatment group (group C), spinal cord injury but without treatment group (group D) and healthy control group (group E). The recovery of neurological function was evaluated by somatosensory evoked potential (SEP) and pathological examination after surgery. Results In group A, in some rats an escaping response in right hind leg occurred, but in other groups, the motor function was not significantly improved. Histological examination showed that transplanted olfactory lamina propria survived in the transplantation area and expanded on certain routes. NF positive nerve fibers passed through the transplantation area. Compared with group B, C, D, the N1-wave latency was(4.71±0. 72)ms 4 weeks after operation(P<0. 01), and the NF density was(7. 31±0. 26) ×104/mm28 weeks after operation in group A(P<0. 05). Conclusions Olfactory lamina propria (OLP) transplantation and ganglioside GM1 treatment have a synergistic effect on SCI.

Objective To establish and evaluate the new method of 8-color flow cytometry (8c-FCM) with two tube detecting bone marrow minimal residual disease (MRD) in B lineage acute lymphoblastic leukemia (B-ALL).MethodsThe MRD cells were analyzed by using two combinations of 8c-FCM antibody panels,gating with CD19/Side scatter(SSC),CD45/CD10 and CD34(or cTdT).Nalm-6 cell of B-ALL was mixed into normal marrow cells,with proportion of 10.00％,1.00％,0.10％,0.01％and 0.005％,and recovery test and reproducibility test were carried with 8c-FCM established to value its accuracy and precision of.Fluorescence intensity was detected on different time points after marked Nalm-6 by antibodies to evaluate the fluorescent stability of the antibodies.The immunophenotyping was analyzed in 39 bone marrow specimens,including 9 cases of normal control,9 cases of B-ALL primary or recurrent,21 cases of complete remission (CR) after chemotherapy or bone marrow transplantation ( BMT),to evaluate the detection of normal B lymphocyte lineage,leukemia cells of B-ALL,MRD cells of B-ALL using the 8c-FCM and compare it with 4c-FCM in being.ResultsThe method of two tube 8c-FCM with main antibodies of CD19,CD45 and CD10 to detect MRD of B-ALL was founded; CV ＜ 2.5％,average recovery rate was 95.81％,when the actual percent of Nalm-6 mixed into normal bone marrow≥0.10％,and the percent of Nalm-6 detected and actual was linear dependent ( r =0.99,P ＜ 0.05 ) ranged from 10.00％ - 0.01 ％,when 106 cells were acquired ; the sensitivity of the method established could reach 0.01％.The fluorescent intensity decreased along with the time after Nalm-6 cell marked,but less than 10％ in 24 hours.Using the antibody combinations and analyze strategy,the immunophenotye of B lymphocyte in normal bone marrow presented four sequential stages:Stage Ⅰ CD45low/CD10stro/CD20 -/CD38stro/CD34 + or cTdT +,Stage ⅡCD45 +/CD10 + / CD20 -/CD38stro/CD34+ or cTdT +,Stage Ⅲ CD45 +/CD10 +/CD20 -/CD38stro/CD34 -or cTdT-,Stage Ⅳ CD45stro/CD10 -/CD20 +/CD38low/CD34or cTdT.Antigen expressions of leukeamic cells of B-ALL primary or recurrent were different compared with control team; there were 5 cases with MRD positive in CR team,and the main antigen expression was consistent with the results from 4c-FCM.The range of the percent of MRD cells was 0.02％ - 5.42％ of the 5 cases of MRD positive.ConclusionsThe new method of two tube 8c-FCM established shows good reproducibility and high accuracy,and can identify normal B lymphocyte populations in bone marrow and regenerated B-precursors in CR cases with MRD cells;compared with 4c-FCM,the new method of two tube 8c-FCM with the fewer specimen is faster and efficient to diagnose MRD of B-ALL.

Objective To observe the effects of transplantation of bone marrow stem cells(BMSCs)transfected with bone morphogenetic protein-2(BMP-2)on fracture healing in rats with diabetes so as to provide a new therapy for diabetic fractures.Methods Fifty male adult Wistar rats aged six weeks randomized to the control group and experimental group were all employed to establish models with diabetic fractures.Under high glucose condition,BMSCs were transfected with BMP-2 by adenovirus vector in vitro.BMSCs transfected by BMP-2 were transplanted into the fracture area of rats in the experimental group,while non-transfected BMSCs into the corresponding area of rats in the control group.X-ray examination was performed at 1,2,3,4 and 6 weeks after transplantation.Bony calluses were collected for HE staining and gray scales of BMP-2 in calluses were determined by immunohistochemical method.Meanwhile,serum levels of BMP-2 were measured by ELtSA.Results The gray scales of BMP-2 in the calluses were 83±3 in the experimental group and 118±4 in the control group at four weeks(P＜0.01).The serum concentrations of BMP-2 were(203.80±8.96)ng/L in the experimental group and(139.15±4.19)ng/L in the control group at four weeks(P＜0.01).Conclusion Transplantation of BMSCs transfected by BMP-2 promotes fracture healing in diabetic rats.

BackgroundReactive oxygen intermediate products lead to the oxidative injury of cells.Retinal pigment epithelial(RPE) cells produce lots of reactive oxygen intermediate products during the swallow of out disc,but how this procedure cause the persistent oxidative injury of RPE cells is poorly understood.Objective The present study was to evaluate the effect of non-lethal H2 O2 -induced persistent oxidative injury on RPE barrier in vitro.MethodsARPE-19 cell links were inoculated on 96 well plate at the density of 8×104 cells/L and the cell climbing slice of 24 well at the density of 4× 104 cells/L.The cells were cultured in DMEM/F12 medium,and the cells cultured for 24 hours in free-serum medium were used in the experiment.0-0.6 mmol/L of H2O2 were added into the medium.Cellular viability was assessed using 3- ( 4,5-dimethylthiazol-2-yl ) -5- ( 3-carboxymethoxyphenyl ) -2- ( 4-sulfophenyl ) 2H-tetrazolium(MTS) assays.Transepithelial electrical resistance (TER) was used to detect cell monolayer forming time after cultureinTrsnswellchamber.Thepermeabilityof cellmonolayer was examinedbyrhodamine isothiocyanate-dextran transepithelial flux,and immunofluorescence was used to investigate the distribution of the junction protein zonula occludens (ZO-1).ResultsThe total difference was found in the cell vitality(A490) among the different concentrations of H2 O2 ( F =991.501,P =0.000 ).Compared with 0 mmoL/L H2 O2 group,the A490 values was gradually lowed from 0.20 mmol/L H2O2 group to 0.60 mmol/L H2O2 group (P ＜ 0.05 ).H2O2 at the concentrations of ＞0.20 mmol/L lowed the viability of RPE cells.The TER value was ( 24.9 ± 1.3 ) Ω · cm2 in 11 days,( 17.8± 1.4)Ω · cm2 in 7 days after inoculation on transwell chamber,showing a significant difference between them (t=5.228,P=0.014).RPE formed the stable tight junction on day 15 with the TER value (25.9±0.9 ) Ω · cm2.The leakage amount ( relative fluorescence intensity ) of the dextran was 255.39 ± 16.44 in non-H2 O2 control group,exhibiting a significant lowing in comparison with free-cell blank group (433.08±51.53)( t =12.515,P =0.006 ),and that of H2 O2 group was significant increased in comparison with non-H2 O2 control group ( t =14.412,P=0.005).Immunofluyorescence assay showed intact intercellular ZO-1 junction in non-H2O2 control group,but the breakage of ZO-1 junction was seen in H2O2 group.ConclusionsThe results indicate that non-lethal H2O2 can destroy RPE barrier and further lead to the persistent oxidative injury of RPE cells.

To investigate the effects of Losartan,an angiotensinⅡ(AngⅡ)receptor(AT_1) antagonist,on pancreatic stellate cells(PSCs)and its possible mechanisms.Methods (1)PSCs were isolated from pancreatic cancerous samples to test the expressions of AT_1 and collagenⅠafter incubated with AngⅡor/and Losartan.(2)Ninety S-D rats were divided into normal group,control group and treatment group,with 30 rats in each.The rats in control and treatment groups were induced pancreatic fibrosis by injection of 2% trinitrobenenze sulfonic acid(TNBS)into biliopancreatic duct.Rats in treat- ment group were then treated with Losartan by garage daily and rats in control group were only given distilled water.The rats were sacrificed on day 3,7,14,21 and 28,respectively,and pancreas were removed.The histological abnormalities were observed by electron microscope.The mRNAs of trans- forming growth factor?_1(TGF?_1)and procollagenⅠwere detected by reverse transcription-polymerase chain reaction(RT-PCR).The expression of TGF?_1 and?-smooth muscle actin(?-SMA)proteins was assessed by immunohistochemistry and the level of?-SMA protein was quantified by Western blot. Results In vitro,there existed AT_1 expression in PSCs,and Losartan reduced expression of collagenⅠ.Losartan treatment reversed the histological abnormalities observed by electron microscope,com- pared to treatment with distill water.The expression of?-SMA,TGF?_1 and procollagenⅠwere signifi- cantly higher in the control group than those in normal group and were reduced by Losartan to different extent in treatment group.Conclusion AT_1 antagonist can inhibit the activation and the profibrogenic action of PSCs by blocking AT_1 receptor-mediated pathways.

DNA Methylation ; Glioma ; metabolism ; Humans ; O(6)-Methylguanine-DNA Methyltransferase ; genetics ; Polymerase Chain Reaction ; methods ; Promoter Regions, Genetic ; genetics

The response rules of pressing pain on the back section in the Governor Vessel in patients with gastro-esophageal reflux disease (GERD) were studied to provide references for the diagnosis and treatment of GERD. Seventy-six cases of GERD were included into an observation group while 30 healthy volunteers were recruited into a control group. A mechanical measurement device of pressing pain that could measure the pain threshold was adapted to observe the pressing pain on the back section in the Governor Vessel in GERD patients and healthy volunteers. The test area is from spinous process of the 1st thoracic vertebra to that of the 12th thoracic vertebra (T1 -T12), including acupoints and non-acupoints on the Governor Vessel. As a result, in the observation group the pain threshold of T5-T7 spinous process clearance, which was the location of Shendao (GV 11), Lingtai (GV 10) and Zhiyang (GV 9), was lower than that in the control group (all P < 0.05). This result indicated that there was significant pressing pain in T5-T7 spinous process clearance in patients with GERD, which could be taken as an important auxiliary diagnosis and a new thinking method in the treatment of GERD with acupuncture.
Adolescent ; Adult ; Aged ; Diagnosis, Differential ; Female ; Gastroesophageal Reflux ; diagnosis ; physiopathology ; Humans ; Male ; Meridians ; Middle Aged ; Pressure ; Sensation ; Thoracic Vertebrae ; physiopathology ; Young Adult

China ; Clinical Trials as Topic ; Drugs, Chinese Herbal ; adverse effects ; therapeutic use ; Humans ; Kidney Diseases ; drug therapy ; Prospective Studies ; Retrospective Studies ; Tripterygium

Australia ; Environmental Monitoring ; Humans ; Occupational Health ; Safety Management ; methods ; organization & administration

OBJECTIVETo investigate the distribution characters and linkage disequilibrium of single nucleotide polymorphisms (SNPs) -148C/T, -455G/A and -854G/A in the promoter region of fibrinogen B(FGB) beta gene.

METHODSGenotype and allele frequencies of FGB beta gene were examined by polymerase chain reaction-restriction fragment length polymorphism and nucleotide sequencing methods in 377 Chinese southern Han individuals. Three FGB beta SNPs Hardy-Weinberg equilibrium and linkage disequilibrium were analyzed with population genetics methods.

RESULTSThe allele frequencies of 3 SNPs are in good agreement with Hardy-Weinberg equilibrium. A total of 9 genotypes among the 377 individuals were identified in 3 SNPs. The genotype frequencies of -148CC, CT and TT were 0.597, 0.358 and 0.045, respectively; the -455G/A genotype frequencies were the same as that of -148C/T SNP; the genotype frequencies of -854GG,GA, AA were 0.820,0.178,0.002, respectively. The frequencies of rare allele -148T, -455A and -854A were 0.224,0.224 and 0.092, respectively, while the common allele frequencies were 0.776 for -148C, 0.776 for -455G, and 0.908 for -854G. There were no statistically significant differences in genotype and allele frequencies between the male and female groups (P>0.05). The relationship between -455G and -148C was completely concordant, but there was a random distribution between -854 and -148 (-445) SNPs.

CONCLUSIONThe results show there is a complete linkage disequilibrium between -148C/T and -455G/A and a negative linkage disequilibrium between -854G/A and -148C/T, as well as between -854G/A and -455G/A. This study has provided population genetics data on FGB beta gene promoter in Chinese southern Han population.

Adult ; Aged ; Aged, 80 and over ; China ; ethnology ; Female ; Fibrinogen ; genetics ; Humans ; Linkage Disequilibrium ; Male ; Middle Aged ; Polymerase Chain Reaction ; Polymorphism, Restriction Fragment Length ; Polymorphism, Single Nucleotide ; Promoter Regions, Genetic